| Chapter title |
Measurement of Chaperone-Mediated Effects on Polyglutamine Protein Aggregation by the Filter Trap Assay
|
|---|---|
| Chapter number | 5 |
| Book title |
Chaperones
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7477-1_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7476-4, 978-1-4939-7477-1
|
| Authors |
Maria A. W. H. van Waarde-Verhagen, Harm H. Kampinga, Waarde-Verhagen, Maria A. W. H., Kampinga, Harm H. |
| Abstract |
The formation of aggregates by polyglutamine-containing (polyQ) proteins in neurons is a key to the pathogenesis of several progressive neurodegenerative diseases such as Huntington's disease (HD) spinocerebellar ataxias (SCAs), and spinal and bulbar muscular atrophy (SBMA). In order to study whether the members of the heat shock protein (HSP) families, by virtue of their molecular chaperone activity, can inhibit the formation of polyQ aggregates, we developed a cell culture model expressing the GFP tagged fragment of exon1 of the huntingtin gene with an expanded polyQ chain and tetracycline inducible chaperones. Expression of mutated Huntington's protein leads to the formation of 2% SDS insoluble high molecular weight polyQ aggregates that are retarded on a cellulose acetate membrane in the so-called filter trap assay (FTA). This chapter explains in detail the protocols of the FTA and how it can be a useful tool to study the effect of HSPs or their functional mutants on aggregation of polyglutamine proteins. Moreover, the assay is useful to investigate how externally added polyQ peptides can act as nucleation seeds for internally expressed polyQ proteins. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 26 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 5 | 19% |
| Student > Ph. D. Student | 4 | 15% |
| Researcher | 3 | 12% |
| Student > Bachelor | 2 | 8% |
| Student > Doctoral Student | 2 | 8% |
| Other | 6 | 23% |
| Unknown | 4 | 15% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 7 | 27% |
| Neuroscience | 6 | 23% |
| Agricultural and Biological Sciences | 3 | 12% |
| Business, Management and Accounting | 1 | 4% |
| Nursing and Health Professions | 1 | 4% |
| Other | 4 | 15% |
| Unknown | 4 | 15% |